is the known causative agent of both cutaneous (CL) and visceral leishmaniasis in Sri Lanka.
CL is considered to be under-reported partly due to relatively poor sensitivity and specificity of microscopic diagnosis.
We compared robustness of three previously described polymerase chain reaction (PCR) based methods to detect
DNA in 38 punch biopsy samples from patients presented with suspected lesions in 2010. Both, Leishmania
genus-specific JW11/JW12 KDNA and LITSR/L5.8S internal transcribed spacer (ITS)1 PCR assays detected
92% (35/38) of the samples whereas a KDNA assay specific for L. donovani
(LdF/LdR) detected only 71% (27/38) of
samples. All positive samples showed a L. donovani
banding pattern upon HaeIII ITS1 PCR-restriction fragment
length polymorphism analysis. PCR assay specificity was evaluated in samples containing Mycobacterium tuberculosis
, Mycobacterium leprae
, and human DNA, and there was no cross-amplification in JW11/JW12 and LITSR/L5.8S
PCR assays. The LdF/LdR PCR assay did not amplify M. leprae
or human DNA although 500 bp and 700 bp bands
were observed in M. tuberculosis
samples. In conclusion, it was successfully shown in this study that it is possible to
diagnose Sri Lankan CL with high accuracy, to genus and species identification, using Leishmania
DNA PCR assays.